Using conditional VgluT2-KO techniques, we found that deletion of VgluT2 in a subpopulation of DA neurons abolished glutamate release from these cells, causing a reduction in BDNF and TrkB expression in midbrain DA neurons and an increase in vulnerability to MPTP-induced DA cell death and locomotor impairment. Restoration of VgluT2 expression in VgluT2-cKO mice normalized BDNF/TrkB expression and attenuated MPTP-induced toxicity in DA neurons and locomotor dysfunction. These findings suggest that reduced VgluT2 expression in DA neurons may be a newly-identified risk factor in the development of neurodegenerative diseases, such as PD, and normalization of VgluT2 expression in DA neurons may be useful in preventing and treating PD or other neurodegenerative diseases.